Tumor Necrosis Factor-.alpha. (TNF-.alpha.) and Lymphotoxin or TNF-.beta. (hereinafter, TNF refers to both TNF-.alpha. and TNF-.beta.) are cytokines which have many effects on cells (Wallach, D. (1986) in: Interferon 7 (Ion Gresser, Ed.), pp. 83-122, Academic Press, London, and Beutler, B. and Cerami, A. (1987) New England J. Med. 316: 379-385). Both TNF-.alpha. and TNF-.beta. initiate their effects by binding to specific cell surface receptors. Some of the effects are likely to be beneficial to the organism: they may destroy, for example, tumor cells or virus infected cells and augment antibacterial activities of granulocytes. But, quite clearly, both TNF-.alpha. and TNF-.beta. have also effects which can be extensively deleterious. There is evidence that over production of TNF-.alpha. can play a major pathogenic role in several diseases. Thus effects of TNF-.alpha., primarily on the vasculature, are now known to be a major cause for symptoms of septic shock (Tracey, K. J. et al. (1986) Science 234: 470-474). In some diseases, TNF may cause excessive loss of weight (cachexia) by suppressing activities of adipocytes and by causing anorexia and TNF-.alpha. was thus called cachectin. It was also described as a mediator of the damage to tissues in rheumatic disease (Beutler, op.cit.) and as a major mediator of the damage observed in graft-versus-host reactions.
There is therefore a necessity in finding out ways to eliminate or antagonize endogenously formed or exogenously administered TNF. Our first attempt in this direction was the development of monoclonal antibodies which neutralize the TNF-.alpha. cytotoxic activity and were shown to protect mice against the lethal effect of TNF-.alpha. under conditions mimicking elicitation of septic shock (as described in our U.S. patent application Ser. No. 06/808,262 of Dec. 12, 1985). However, therapy with murine monoclonal antibodies, especially if administered repetitively, may not always be advisable in humans. Therefore the need was felt for development of biological agents which could similarly antagonize the deleterious effects of TNF.
Prior to the filing date of the priority application of the present application, there was no information as to the existence of biological agents which could antagonize the cytotoxic activity of THF. There were publications describing uromodulin, a 85-kDa immunosuppressive glycoprotein isolated from the urine of pregnant women (Muchmore, Andrew V. and Decker, Jean M. (1985) Science 229:479-481), that was shown to be a high affinity ligand for and a potent inhibitor of interleukin 1 (IL-1) (Muchmore, Andrew V. and Decker, Jean M. (1986) J. Biol. Chem. 261:13404-13407; Brown, K. M. et al. (1986) Proc. Natl. Acad. Sci. U.S.A. 83:9119-9123). Uromodulin was later shown to be identical to the Tamm-Horsfall glycoprotein, the most abundant protein of renal origin in normal urine (Pennica, Diane et al. (1987) Science 236:83-88). Another inhibitor of IL-1 found in the urine of febrile patients was disclosed in some publications (Liao, Zenghua et al. (1984) J. Exp. Med. 159:126-136; Seckinger, Phillippe et al. (1987) J. Immunol. 139:1546-1549). It was shown that this urine inhibitor of IL-1 affects numerous biological activities of both forms of recombinant IL-1, IL-1.alpha. and IL-1.beta. to the same extent. Although human TNF-.alpha. shares some of the biological activities of IL-1, this IL-1 inhibitor did not inhibit the biological activities of TNF-.alpha. (Seckinger, Phillippe et al. (1987) J. Immunol. 139:1541-1545).
Subsequent to the filing date of the priority application of the present application, it was disclosed that uromodulin and the Tamm-Horsfall glycoprotein bind recombinant IL-1.alpha., IL-1.beta. and TNF-.alpha. in a lectin-like interaction and it was suggested that it may play an important role in the regulation of circulating levels of these lymphokines (Hession, Catherine et al. (1987) Science 237:1479-1484). Although uromodulin does not inhibit the cytotoxic activity of TNF-.alpha. as monitored by lysis of tumor cell targets, it interacts with recombinant TNF-.alpha. via carbohydrate chains and this interaction may be critical in promoting clearance and/or reducing in vivo toxicity of TNF and other lympyhokines (Sherblom, Anne P. (1988) J. Biol. Chem. 263:5418-5424). In a recent publication by Seckinger et al. (J. Exp. Med. (1988) 167:1511-1516) a human inhibitor of TNF-.alpha. obtained from the urine of febrile patients was described as a 40-60 Kda protein inhibiting the cytotoxic activity of TNF-.alpha.. It was shown to differ from uromodulin and from the above-mentioned IL-1 inhibitor.